The Role of Emission Size Distribution on the Efficacy of New Technologies to Reduce Methane Emissions from the Oil and Gas Sector.

Methane emissions from oil and gas operations exhibit skewed distributions. New technologies such as aerial-based leak detection surveys promise cost-effective detection of large emitters (greater than 10 kg/h). Recent policies such as the US Environmental Protection Agency (EPA) methane rule that allow the use of new technologies as part of leak detection and repair (LDAR) programs require a demonstration of equivalence with existing optical gas imaging (OGI) based LDAR programs. In this work, we illustrate the impact of emission size distribution on the equivalency condition between the OGI and site-wide survey technologies. Emission size distributions compiled from aerial measurements include significantly more emitters between 1 and 10 kg/h and lower average emission rates for large emitters compared to the emission distribution in the EPA rule. As a result, we find that equivalence may be achieved at lower site-wide survey frequencies when using technologies with detection thresholds below 10 kg/h, compared to the EPA rule. However, equivalence cannot be achieved with a detection threshold of 30 kg/h at any survey frequency, because most emitters across most US basins exhibit emission rates below 30 kg/h. We find that equivalence is a complex tradeoff among technology choice, design of LDAR programs, and survey frequency that can have more than one unique solution set.

Beyond Death: Funeral Rituals and Megalithic Traditions Among the Bhil Tribes of India.

The earliest traces of funeral rituals in the Indian Subcontinent date back to the Mesolithic period. Data from various parts of the country clearly indicate that this tradition gained popularity during the Neolithic and Iron Age periods in India. Besides, during the Iron-age period we found evidence of megalithic tradition, and the megalithic sites found in the Indian subcontinent reveal an elaborate process of death rituals. This present study focuses on first-hand data obtained from the south-eastern part of Rajasthan, India. The data unveils many hidden aspects of funeral rituals and megalithic traditions related to the Bhils community. To gain a better understanding of death rituals, beliefs about afterlife, and the erection of memorial pillars on behalf of the deceased, the author compares the death rituals of the Bhils with those of the Munda communities in central eastern parts of India.

The leucine-responsive regulatory proteins/feast-famine regulatory proteins: an ancient and complex class of transcriptional regulators in bacteria and archaea.

Since the discovery of the Escherichia coli leucine-responsive regulatory protein (Lrp) almost 50 years ago, hundreds of Lrp homologs have been discovered, occurring in 45% of sequenced bacteria and almost all sequenced archaea. Lrp-like proteins are often referred to as the feast/famine regulatory proteins (FFRPs), reflecting their common regulatory roles. Acting as either global or local transcriptional regulators, FFRPs detect the environmental nutritional status by sensing small effector molecules (usually amino acids) and regulate the expression of genes involved in metabolism, virulence, motility, nutrient transport, stress tolerance, and antibiotic resistance to implement appropriate behaviors for the specific ecological niche of each organism. Despite FFRPs' complexity, a significant role in gene regulation, and prevalence throughout prokaryotes, the last comprehensive review on this family of proteins was published about a decade ago. In this review, we integrate recent notable findings regarding E. coli Lrp and other FFRPs across bacteria and archaea with previous observations to synthesize a more complete view on the mechanistic details and biological roles of this ancient class of transcription factors.

Resilience and robustness of alphaproteobacterial methanotrophs upon methane feast-famine scenarios.

Most of methane (CH4) emissions contain low CH4 concentrations and typically occur at irregular intervals, which hinders the implementation and performance of methane abatement processes. This study aimed at understanding the metabolic mechanisms that allow methane oxidizing bacteria (MOB) to survive for long periods of time under methane starvation. To this aim, we used an omics-approach and studied the diversity and metabolism of MOB and non-MOB in bioreactors exposed to low CH4 concentrations under feast-famine cycles of 5 days and supplied with nutrient-rich broth. The 16S rRNA and the pmoA transcripts revealed that the most abundant and active MOB during feast and famine conditions belonged to the alphaproteobacterial genus Methylocystis (91-65%). The closest Methylocystis species were M. parvus and M. echinoides. Nitrifiers and denitrifiers were the most representative non-MOB communities, which likely acted as detoxifiers of the system. During starvation periods, the induced activity of CH4 oxidation was not lost, with the particulate methane monooxygenase of alphaproteobacterial MOB playing a key role in energy production. The polyhydroxyalkanoate and nitrification metabolisms of MOB had also an important role during feast-famine cycles, maintaining cell viability when CH4 concentrations were negligible. This research shows that there is an emergence and resilience of conventional alphaproteobacterial MOB, being the genus Methylocystis a centrepiece in environments exposed to dilute and intermittent methane emissions. This knowledge can be applied to the operation of bioreactors subjected to the treatment of dilute and discontinuous emissions via controlled bioaugmentation.

New Technologies Can Cost Effectively Reduce Oil and Gas Methane Emissions, but Policies Will Require Careful Design to Establish Mitigation Equivalence.

Reducing methane emissions from oil and gas systems is a central component of US and international climate policy. Leak detection and repair (LDAR) programs using optical gas imaging (OGI)-based surveys are routinely used to mitigate fugitive emissions or leaks. Recently, new technologies and platforms such as planes, drones, and satellites promise more cost-effective mitigation than existing approaches. To be approved for use in LDAR programs, new technologies must demonstrate emissions mitigation equivalent to existing approaches. In this work, we use the FEAST modeling tool to (a) identify cost vs mitigation trade-offs that arise from using new technologies and (b) provide a framework for effective design of alternative LDAR programs. We identify several critical insights. First, LDAR programs can trade sensitivity for speed without sacrificing mitigation outcomes. Second, low sensitivity or high detection threshold technologies have an effective upper bound on achievable mitigation that is independent of the survey frequency. Third, the cost effectiveness of tiered LDAR programs using site-level detection technologies depends on their ability to distinguish leaks from routine venting. Finally, "technology equivalence" based on mitigation outcomes differs across basins and should be evaluated independently. The FEAST model will enable operators and regulators to systematically evaluate new technologies in next-generation LDAR programs.

Fluctuations in glucose availability prevent global proteome changes and physiological transition during prolonged chemostat cultivations of Saccharomyces cerevisiae.

Chemostat cultivation mode imposes selective pressure on the cells, which may result in slow adaptation in the physiological state over time. We applied a two-compartment scale-down chemostat system imposing feast-famine conditions to characterize the long-term (100 s of hours) response of Saccharomyces cerevisiae to fluctuating glucose availability. A wild-type strain and a recombinant strain, expressing an insulin precursor, were cultured in the scale-down system, and analyzed at the physiological and proteomic level. Phenotypes of both strains were compared with those observed in a well-mixed chemostat. Our results show that S. cerevisiae subjected to long-term chemostat conditions undergoes a global reproducible shift in its cellular state and that this transition occurs faster and is larger in magnitude for the recombinant strain including a significant decrease in the expression of the insulin product. We find that the transition can be completely avoided in the presence of fluctuations in glucose availability as the strains subjected to feast-famine conditions under otherwise constant culture conditions exhibited constant levels of the measured proteome for over 250 hr. We hypothesize possible mechanisms responsible for the observed phenotypes and suggest experiments that could be used to test these mechanisms.

Escherichia coli metabolism under short-term repetitive substrate dynamics: adaptation and trade-offs.

BACKGROUND: Microbial metabolism is highly dependent on the environmental conditions. Especially, the substrate concentration, as well as oxygen availability, determine the metabolic rates. In large-scale bioreactors, microorganisms encounter dynamic conditions in substrate and oxygen availability (mixing limitations), which influence their metabolism and subsequently their physiology. Earlier, single substrate pulse experiments were not able to explain the observed physiological changes generated under large-scale industrial fermentation conditions. RESULTS: In this study we applied a repetitive feast-famine regime in an aerobic Escherichia coli culture in a time-scale of seconds. The regime was applied for several generations, allowing cells to adapt to the (repetitive) dynamic environment. The observed response was highly reproducible over the cycles, indicating that cells were indeed fully adapted to the regime. We observed an increase of the specific substrate and oxygen consumption (average) rates during the feast-famine regime, compared to a steady-state (chemostat) reference environment. The increased rates at same (average) growth rate led to a reduced biomass yield (30% lower). Interestingly, this drop was not followed by increased by-product formation, pointing to the existence of energy-spilling reactions. During the feast-famine cycle, the cells rapidly increased their uptake rate. Within 10 s after the beginning of the feeding, the substrate uptake rate was higher (4.68 µmol/gCDW/s) than reported during batch growth (3.3 µmol/gCDW/s). The high uptake led to an accumulation of several intracellular metabolites, during the feast phase, accounting for up to 34% of the carbon supplied. Although the metabolite concentrations changed rapidly, the cellular energy charge remained unaffected, suggesting well-controlled balance between ATP producing and ATP consuming reactions. CONCLUSIONS: The adaptation of the physiology and metabolism of E. coli under substrate dynamics, representative for large-scale fermenters, revealed the existence of several cellular mechanisms coping with stress. Changes in the substrate uptake system, storage potential and energy-spilling processes resulted to be of great importance. These metabolic strategies consist a meaningful step to further tackle reduced microbial performance, observed under large-scale cultivations.

Malaria and acute kidney injury.

Malaria is a parasitic infection transmitted by mosquitos, resulting in significant morbidity and mortality. It affects 212 million worldwide, causing death in up to 303,000 children annually. In the USA, up to 1700 people are affected yearly. Although the prevalence in developed countries is less than in developing countries, travelers from low transmission areas, and those from endemic areas who later return, are very susceptible to malaria and its complications. Severe malaria can cause significant multiorgan dysfunction including acute kidney injury (AKI). The pathogenesis is not clearly understood but proposed mechanisms include acute tubular necrosis (ATN) due to impediments in renal microcirculation, infection-triggered proinflammatory reactions within the kidney, and metabolic disturbances. Providers must consider malarial infection in cases of AKI in someone with a travel history, as early recognition and treatment are crucial to improving outcomes. This article will review malaria-induced AKI in order to provide a better understanding of this infection's effect on the kidneys.

Event-Based Feature Extraction Using Adaptive Selection Thresholds.

Unsupervised feature extraction algorithms form one of the most important building blocks in machine learning systems. These algorithms are often adapted to the event-based domain to perform online learning in neuromorphic hardware. However, not designed for the purpose, such algorithms typically require significant simplification during implementation to meet hardware constraints, creating trade offs with performance. Furthermore, conventional feature extraction algorithms are not designed to generate useful intermediary signals which are valuable only in the context of neuromorphic hardware limitations. In this work a novel event-based feature extraction method is proposed that focuses on these issues. The algorithm operates via simple adaptive selection thresholds which allow a simpler implementation of network homeostasis than previous works by trading off a small amount of information loss in the form of missed events that fall outside the selection thresholds. The behavior of the selection thresholds and the output of the network as a whole are shown to provide uniquely useful signals indicating network weight convergence without the need to access network weights. A novel heuristic method for network size selection is proposed which makes use of noise events and their feature representations. The use of selection thresholds is shown to produce network activation patterns that predict classification accuracy allowing rapid evaluation and optimization of system parameters without the need to run back-end classifiers. The feature extraction method is tested on both the N-MNIST (Neuromorphic-MNIST) benchmarking dataset and a dataset of airplanes passing through the field of view. Multiple configurations with different classifiers are tested with the results quantifying the resultant performance gains at each processing stage.

An evaluation of the mourning tradition, the "First Feast," in the context of palliative care: The possibility of incorporating cultural rituals into palliative care.

OBJECTIVE: Grief is intertwined with cultural and religious rituals that are highly appreciated in the quality standards of palliative care. Here, we aimed to investigate whether a cultural mourning ritual, the "First Feast," can be used by palliative care teams to ease the grief response of the deceased patient's relatives. METHOD: A questionnaire with 23 questions about the prevalence of the First Feast tradition, the content, the pros and cons, and whether it would be useful for the grieving relatives of deceased patients was prepared and given to the palliative care patients' relatives. The data were evaluated using the chi-square test. RESULT: A total of 427 participants were enrolled in the study; 60.7% were female and the mean age was 36 (±13.4). A total of 76.8% of the participants were from the Tokat region and 77.8% (n = 332) performed the First Feast tradition. A significant difference was observed among participants with Tokat origins and non-Tokat origins in terms of awareness of the tradition (84.8% and 69.7%, respectively; p = 0.001). Ninety-one percent of the participants acknowledged that the tradition helped to ease the grief response of the relatives. SIGNIFICANCE OF RESULTS: The First Feast, a mourning tradition performed in Tokat and other parts of Turkey, might be a useful auxiliary method for palliative care teams to help grieving families.

Selecting optimal feast-to-famine ratio for a new polyhydroxyalkanoate (PHA) production system fed by valerate-dominant sludge hydrolysate.

The feast-to-famine ratio (F/F) represents the extent of selective pressure during polyhydroxyalkanoate (PHA) culture selection. This study evaluated the effects of F/F on a new PHA production system by an enriched culture with valerate-dominant sludge hydrolysate and selected the optimal F/F. After the original F/F 1/3 was modified to 1/1, 1/2, 1/4, and 1/5, F/F did not affect their lengths of feast phase, but affected their biomass growth behaviors during the famine phase and PHA-producing abilities. The optimal F/F was 1/2, and compared with 1/3, it increased the maximal PHA content and the fraction of 3-hydroxyvalerate (3HV) and 3-hydroxy-2-methylvalerate (3H2MV) monomers, with higher productivity and better polymer properties. Although F/F 1/2 impaired the advantage of the dominant genus Delftia, it improved the PHA production rate while decreased biomass growth rate, meanwhile enhancing the utilization and conversion of valerate. These findings indicate that in contrast to previous studies using acetate-dominant substrate for PHA production, the new system fed by valerate-dominant substrate can adopt a higher F/F.

Crystal Structure of Mycobacterium tuberculosis H37Rv AldR (Rv2779c), a Regulator of the ald Gene: DNA BINDING AND IDENTIFICATION OF SMALL MOLECULE INHIBITORS.

Here we report the crystal structure of M. tuberculosis AldR (Rv2779c) showing that the N-terminal DNA-binding domains are swapped, forming a dimer, and four dimers are assembled into an octamer through crystal symmetry. The C-terminal domain is involved in oligomeric interactions that stabilize the oligomer, and it contains the effector-binding sites. The latter sites are 30-60% larger compared with homologs like MtbFFRP (Rv3291c) and can consequently accommodate larger molecules. MtbAldR binds to the region upstream to the ald gene that is highly up-regulated in nutrient-starved tuberculosis models and codes for l-alanine dehydrogenase (MtbAld; Rv2780). Further, the MtbAldR-DNA complex is inhibited upon binding of Ala, Tyr, Trp and Asp to the protein. Studies involving a ligand-binding site G131T mutant show that the mutant forms a DNA complex that cannot be inhibited by adding the amino acids. Comparative studies suggest that binding of the amino acids changes the relative spatial disposition of the DNA-binding domains and thereby disrupt the protein-DNA complex. Finally, we identified small molecules, including a tetrahydroquinoline carbonitrile derivative (S010-0261), that inhibit the MtbAldR-DNA complex. The latter molecules represent the very first inhibitors of a feast/famine regulatory protein from any source and set the stage for exploring MtbAldR as a potential anti-tuberculosis target.

A 9-pool metabolic structured kinetic model describing days to seconds dynamics of growth and product formation by Penicillium chrysogenum.

A powerful approach for the optimization of industrial bioprocesses is to perform detailed simulations integrating large-scale computational fluid dynamics (CFD) and cellular reaction dynamics (CRD). However, complex metabolic kinetic models containing a large number of equations pose formidable challenges in CFD-CRD coupling and computation time afterward. This necessitates to formulate a relatively simple but yet representative model structure. Such a kinetic model should be able to reproduce metabolic responses for short-term (mixing time scale of tens of seconds) and long-term (fed-batch cultivation of hours/days) dynamics in industrial bioprocesses. In this paper, we used Penicillium chrysogenum as a model system and developed a metabolically structured kinetic model for growth and production. By lumping the most important intracellular metabolites in 5 pools and 4 intracellular enzyme pools, linked by 10 reactions, we succeeded in maintaining the model structure relatively simple, while providing informative insight into the state of the organism. The performance of this 9-pool model was validated with a periodic glucose feast-famine cycle experiment at the minute time scale. Comparison of this model and a reported black box model for this strain shows the necessity of employing a structured model under feast-famine conditions. This proposed model provides deeper insight into the in vivo kinetics and, most importantly, can be straightforwardly integrated into a computational fluid dynamic framework for simulating complete fermentation performance and cell population dynamics in large scale and small scale fermentors. Biotechnol. Bioeng. 2017;114: 1733-1743. © 2017 Wiley Periodicals, Inc.

Community proteomics provides functional insight into polyhydroxyalkanoate production by a mixed microbial culture cultivated on fermented dairy manure.

Polyhydroxyalkanoates (PHAs) are bio-based, biodegradable polyesters that can be produced from organic-rich waste streams using mixed microbial cultures (MMCs). To maximize PHA production, MMCs are enriched for bacteria with a high polymer storage capacity through the application of aerobic dynamic feeding (ADF) in a sequencing batch reactor (SBR), which consequently induces a feast-famine metabolic response. Though the feast-famine response is generally understood empirically at a macro-level, the molecular level is less refined. The objective of this study was to investigate the microbial community composition and proteome profile of an enriched MMC cultivated on fermented dairy manure. The enriched MMC exhibited a feast-famine response and was capable of producing up to 40 % (wt. basis) PHA in a fed-batch reactor. High-throughput 16S rRNA gene sequencing revealed a microbial community dominated by Meganema, a known PHA-producing genus not often observed in high abundance in enrichment SBRs. The application of the proteomic methods two-dimensional electrophoresis and LC-MS/MS revealed PHA synthesis, energy generation, and protein synthesis prominently occurring during the feast phase, corroborating bulk solution variable observations and theoretical expectations. During the famine phase, nutrient transport, acyl-CoA metabolism, additional energy generation, and housekeeping functions were more pronounced, informing previously under-determined MMC functionality under famine conditions. During fed-batch PHA production, acetyl-CoA acetyltransferase and PHA granule-bound phasin proteins were in increased abundance relative to the SBR, supporting the higher PHA content observed. Collectively, the results provide unique microbial community structural and functional insight into feast-famine PHA production from waste feedstocks using MMCs.

Fate of aerobic granular sludge in the long-term: The role of EPSs on the clogging of granular sludge porosity.

This work aims to investigate the stability of aerobic granular sludge in the long term, focusing on the clogging of the granular sludge porosity exerted by the extracellular polymeric substances (EPSs). The effects of different cycle lengths (short and long-term cycle) on the granular sludge stability were investigated. Results obtained outlined that during the short duration cycle, the formation and breakage of the aerobic granules were continuously observed. During this period, the excess of EPS production contributed to the clogging of the granules porosity, causing their breakage in the long run. During the long-duration cycle, the extended famine period entailed a greater EPSs consumption by bacteria, thus limiting the clogging of the porosity, and allowed obtaining stable aerobic granules. Reported results demonstrated that an excess in EPSs content could be detrimental to the stability of aerobic granular sludge in the long-term.

Modeling the competition between PHA-producing and non-PHA-producing bacteria in feast-famine SBR and staged CSTR systems.

Although the enrichment of specialized microbial cultures for the production of polyhydroxyalkanoates (PHA) is generally performed in sequencing batch reactors (SBRs), the required feast-famine conditions can also be established using two or more continuous stirred-tank reactors (CSTRs) in series with partial biomass recirculation. The use of CSTRs offers several advantages, but will result in distributed residence times and a less strict separation between feast and famine conditions. The aim of this study was to investigate the impact of the reactor configuration, and various process and biomass-specific parameters, on the enrichment of PHA-producing bacteria. A set of mathematical models was developed to predict the growth of Plasticicumulans acidivorans-as a model PHA producer-in competition with a non-storing heterotroph. A macroscopic model considering lumped biomass and an agent-based model considering individual cells were created to study the effect of residence time distribution and the resulting distributed bacterial states. The simulations showed that in the 2-stage CSTR system the selective pressure for PHA-producing bacteria is significantly lower than in the SBR, and strongly affected by the chosen feast-famine ratio. This is the result of substrate competition based on both the maximum specific substrate uptake rate and substrate affinity. Although the macroscopic model overestimates the selective pressure in the 2-stage CSTR system, it provides a quick and fairly good impression of the reactor performance and the impact of process and biomass-specific parameters.

Source apportionment based on EEM-PARAFAC combined with microbial tracing model and its implication in complex pollution area, Wujin District, China.

Further improving the quality of surface water is becoming more difficult after the control of main point-sources, especially in the complex pollution area with mixed industrial and agricultural productions, whereas the pollution source apportionment might be the key to quantify different pollution sources and developing some effective measures. In this study, a technical framework for source apportionment based on three-dimensional fluorescence and microbial traceability model is developed. Based on screening of the main environmental factors and their spatiotemporal characteristics, potential pollution sources have been tentatively identified. Then, the pollution sources are further tested based on the analysis of fluorescence excitation-emission matrix (EEM) and the similarity of fluorescence components in surface water and potential pollution sources. At the same time, the correlation between microbial species and pollution sources is constructed by analyzing the spatiotemporal characteristics of microbial composition and the response of main species to environmental factors. Therefore, pollution source apportionment is quantified using PCA-APCS-MLR, Fast Expectation-maximization for Microbial Source Tracking (FEAST), and Bayesian community-wide culture-independent microbial source tracking (SourceTracker). PCA-APCS-MLR could not effectively distinguish the contributions of different industrial sources in the complex environment of this study, and the contribution of unknown sources was high (average 39.60%). In contrast, the microbial traceability model can accurately identify the contribution of 7 pollution sources and natural sources, effectively reduce the proportion of unknown sources (average of FEAST is 19.81%, SourceTracker is 16.72%), and show better pollution identification and distribution capabilities. FEAST exhibits a more sensitive potential in source apportionment and shorter calculation time than SourceTracker, thus might be used to guide the precise regional pollution control, especially in the complex pollution environments.

Phosphorus removal from fermented dairy manure concurrent with polyhydroxybutyrate-co-valerate synthesis under aerobic conditions.

Wastewater phosphorus removal achieved biologically is associated with the process known as enhanced biological phosphorus removal (EBPR). In contrast with canonical EBPR operations that employ alternating anaerobic-aerobic conditions and achieve asynchronous carbon and phosphorus storage, research herein focused on phosphorus removal achieved under aerobic conditions synchronously with volatile fatty acid (VFA) storage as polyhydroxybutyrate-co-valerate (PHBV). 90.3 ± 3.4 % soluble phosphorus removal was achieved from dairy manure fermenter liquor; influent and effluent concentrations were 38.6 ± 9.5 and 3.7 ± 0.8 mgP/L, respectively. Concurrently, PHBV yield ranged from 0.17 to 0.64 mgCOD/mgCOD, yielding 147-535 mgCODPHBV/L. No evidence of EBPR mechanisms was observed, nor were canonical phosphorus accumulating organisms present; additionally, the polyphosphate kinase gene was not present in the microbial biomass. Phosphorus removal was primarily associated with biomass growth and secondarily with biomass complexation. Results demonstrate that concurrent PHBV synthesis and phosphorus recovery can be achieved microbially under aerobic dynamic feeding conditions when fed nutrient rich wastewater.

Feast/famine ratio regulates the succession of heterotrophic nitrification-aerobic denitrification and autotrophic ammonia oxidizing bacteria in halophilic aerobic granular sludge treating saline wastewater.

Heterotrophic nitrification-aerobic denitrification (HN-AD) shows innovation potential of wastewater treatment process in a single tank. However, how to enrich HN-AD bacteria in activated sludge to enhance their contribution remained unknown. This study explored the impact of the feast/famine (F/F) ratio on the succession of autotrophic ammonia oxidizing bacteria (AOB) and HN-AD bacteria in a halophilic aerobic granular sludge (HAGS) system. As the F/F ratio decreased from 1/9 to 1/15, the total inorganic nitrogen (TIN) removal performance significantly decreased. The proportion of heterotrophic bacteria was dropped from 79.0 % to 33 %. Accordingly, the relative abundance of Paracoccus decreased from 70.8 % to 25.4 %, and the copy number of the napA gene was reduced from 2.2 × 1010 copies/g HAGS to 8.1 × 109 copies/g HAGS. It found the F/F ratio regulated the population succession of autotrophic AOB and HN-AD bacteria, thereby providing a solution to achieve the enrichment of HN-AD bacteria in HAGS.

Selective genes expression and metabolites transformation drive a robust nitrite accumulation during nitrate reduction under alternating feast-famine condition.

Nitrite production via denitrification has been regarded as a key approach for survival of anaerobic ammonium oxidation (anammox) bacteria. Despite the important carbon substrate, little is known about the role of differential genes expression and extracellular metabolite regulation among diverse microbial communities. In this study, a novel alternating feast-famine strategy was proposed and demonstrated to efficiently accumulate nitrite in a low-nitrogen loading rate (NLR) (0.2∼0.8 kg N/m3/d) denitrification system. Highly selective expression of denitrifying genes was revealed as key regulators. Interestingly, in absence of carbon source (ACS) condition, the expression of narG and narI/V genes responsible for reduction of nitrate to nitrite jumped to 2.5 and 5.1 times higher than that in presence of carbon source (PCS) condition with carbon to nitrate ratio of 3.0. This fortunately facilitated a rapid nitrite accumulation once acetate was added, despite a significantly down-regulated narG and narI/narV and up-regulated nirS/nirK. This strategy selected Thauera as the most dominant denitrifier (50.2 %) with the highest contribution to narG and narI/narV genes, responsible for the high nitrite accumulation. Additionally, extracellular xylose, pyruvate, and glucose jointly promoted carbon-central metabolic pathway of key denitrifiers in ACS stage, playing an important role in the process of self-growth and selective enrichment of functional bacteria. The relatively rapid establishment and robust performance obtained in this study shows an engineering-feasible and economically-favorable solution for the regulation of partial denitrification in practical application.